In this paper, a novel electrochemical sensor was developed for the rapid detection of G-G mismatched DNA based on hexaammineruthenium(III) chloride ([Ru(NH 3 ) 6 ]C l3 ) as a redox indicator. The sensor platform was constructed by immobilizing small molecules (NC-linker) on the gold electrode via amide bonds. The as-prepared NC-linker as the nucleic acids recognition molecule can interact with the G base of DNA. After the sensor was incubated with G-G mismatched DNA, the double-stranded DNA (dsDNA) acted as carriers of the signal tags-[Ru(NH3)6]Cl3, which resulted in a remarkable electrochemical signal. More binding of [Ru(NH 3 ) 6 ]C l3 led to increases of the electrochemical signal. Other mismatched DNA produced only a low response, as well as complementary DNA. Thus G-G mismatched DNA can be easily discriminated from other mismatched and complementary DNA based on the sensor. Furthermore, the method was simple, rapid and repeatable for the detection of G-G mismatched DNA. The selective detection of target dsDNA was achieved by a relative current ratio of the target and control DNA. These results demonstrated that this strategy could provide great promise for the rapid and specific detection of other sequence-specific DNA.
Hong Guo Chen,Wang Ren,Jing Jia,Ji Feng,Zhong Feng Gao,Nian Bing Li,Hong Qun Luo
We describe here a fluorometric assay for sensitive detection of oligonucleotides , based on a target recycling amplification strategy driven by toehold-mediated strand displacement reaction and on exonuclease III (Exo Ш)-assisted fluorescence background suppression strategy. The network consists of a pair of partially complementary DNA hairpins (HP 1 and HP 2 ) with 3′ overhang ends, between which the spontaneous hybridization is kinetically hindered by the stems. The target DNA is repeatedly used to trigger a recycling progress between the hairpins, generating numerous HP 1 –HP 2 duplex complexes. Exo III was then employed to digest the double strand parts of the residual hairpins and the intermediate products. The fluorescent dye , SYBR Green I , binds to the double-strand DNA products and emits strong fluorescence to achieve sensitive detection of the target DNA with the detection limit of 5.34 pM. Moreover, this proposed strategy showed high discrimination efficiency towards target DNA against mismatched DNA and was successfully applied in the analysis of human serum sample.
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